Broccoli line BRM-53-5943SI

ABSTRACT

The invention provides seed and plants of broccoli hybrid RX 05990035 and the parent lines thereof. The invention thus relates to the plants, seeds and tissue cultures of broccoli hybrid RX 05990035 and the parent lines thereof, and to methods for producing a broccoli plant produced by crossing such plants with themselves or with another broccoli plant, such as a plant of another genotype. The invention further relates to seeds and plants produced by such crossing. The invention further relates to parts of such plants.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the priority of U.S. Provisional Appl. Ser. No.61/593,251, filed Jan. 31, 2012, the entire disclosure of which isincorporated herein by reference.

FIELD OF THE INVENTION

The present invention relates to the field of plant breeding and, morespecifically, to the development of broccoli hybrid RX 05990035 and theparent broccoli lines BRM-53-3936CMS and BRM-53-5943SI.

BACKGROUND OF THE INVENTION

The goal of vegetable breeding is to combine various desirable traits ina single variety/hybrid. Such desirable traits may include any traitdeemed beneficial by a grower and/or consumer, including greater yield,resistance to insects or disease, tolerance to environmental stress, andnutritional value.

Breeding techniques take advantage of a plant's method of pollination.There are two general methods of pollination: a plant self-pollinates ifpollen from one flower is transferred to the same or another flower ofthe same plant or plant variety. A plant cross-pollinates if pollencomes to it from a flower of a different plant variety.

Plants that have been self-pollinated and selected for type over manygenerations become homozygous at almost all gene loci and produce auniform population of true breeding progeny, a homozygous plant. A crossbetween two such homozygous plants of different genotypes produces auniform population of hybrid plants that are heterozygous for many geneloci. Conversely, a cross of two plants each heterozygous at a number ofloci produces a population of hybrid plants that differ genetically andare not uniform. The resulting non-uniformity makes performanceunpredictable.

The development of uniform varieties requires the development ofhomozygous inbred plants, the crossing of these inbred plants, and theevaluation of the crosses. Pedigree breeding and recurrent selection areexamples of breeding methods that have been used to develop inbredplants from breeding populations. Those breeding methods combine thegenetic backgrounds from two or more plants or various other broad-basedsources into breeding pools from which new lines and hybrids derivedtherefrom are developed by selfing and selection of desired phenotypes.The new lines and hybrids are evaluated to determine which of those havecommercial potential.

SUMMARY OF THE INVENTION

In one aspect, the present invention provides a broccoli plant of thehybrid designated RX 05990035 the broccoli line BRM-53-3936CMS orbroccoli line BRM-53-5943SI. Also provided are broccoli plants havingall the physiological and morphological characteristics of such a plant.Parts of these broccoli plants are also provided, for example, includingpollen, an ovule, a floret, a head, and a cell of the plant.

In another aspect of the invention, a plant of broccoli hybrid RX05990035 and/or broccoli lines BRM-53-3936CMS and BRM-53-5943SIcomprising an added heritable trait is provided. The heritable trait maycomprise a genetic locus that is, for example, a dominant or recessiveallele. In one embodiment of the invention, a plant of broccoli hybridRX 05990035 and/or broccoli lines BRM-53-3936CMS and BRM-53-5943SI isdefined as comprising a single locus conversion. In specific embodimentsof the invention, an added genetic locus confers one or more traits suchas, for example, herbicide tolerance, insect resistance, diseaseresistance, and modified carbohydrate metabolism. In furtherembodiments, the trait may be conferred by a naturally occurring geneintroduced into the genome of a line by backcrossing, a natural orinduced mutation, or a transgene introduced through genetictransformation techniques into the plant or a progenitor of any previousgeneration thereof. When introduced through transformation, a geneticlocus may comprise one or more genes integrated at a single chromosomallocation.

The invention also concerns the seed of broccoli hybrid RX 05990035and/or broccoli lines BRM-53-3936CMS and BRM-53-5943SI. The broccoliseed of the invention may be provided, in particular embodiments, as anessentially homogeneous population of broccoli seed of broccoli hybridRX 05990035 and/or broccoli lines BRM-53-3936CMS and BRM-53-5943SI.Essentially homogeneous populations of seed are generally free fromsubstantial numbers of other seed. Therefore, seed of hybrid RX 05990035and/or broccoli lines BRM-53-3936CMS and BRM-53-5943SI may be provided,in certain embodiments of the invention, as forming at least about 97%of the total seed, including at least about 98%, 99% or more of theseed. The seed population may be separately grown to provide anessentially homogeneous population of broccoli plants designated RX05990035 and/or broccoli lines BRM-53-3936CMS and BRM-53-5943SI.

In yet another aspect of the invention, a tissue culture of regenerablecells of a broccoli plant of hybrid RX 05990035 and/or broccoli linesBRM-53-3936CMS and BRM-53-5943SI is provided. The tissue culture willpreferably be capable of regenerating broccoli plants capable ofexpressing all of the physiological and morphological characteristics ofthe starting plant, and of regenerating plants having substantially thesame genotype as the starting plant. Examples of some of thephysiological and morphological characteristics of the hybrid RX05990035 and/or broccoli lines BRM-53-3936CMS and BRM-53-5943SI includethose traits set forth in the tables herein. The regenerable cells insuch tissue cultures may be derived, for example, from embryos,meristems, cotyledons, pollen, leaves, anthers, roots, root tips,pistils, flowers, seed and stalks. Still further, the present inventionprovides broccoli plants regenerated from a tissue culture of theinvention, the plants having all the physiological and morphologicalcharacteristics of hybrid RX 05990035 and/or broccoli linesBRM-53-3936CMS and BRM-53-5943SI.

In still yet another aspect of the invention, processes are provided forproducing broccoli seeds, plants and parts thereof, which processesgenerally comprise crossing a first parent broccoli plant with a secondparent broccoli plant, wherein at least one of the first or secondparent broccoli plants is a plant of broccoli line BRM-53-3936CMS orbroccoli line BRM-53-5943SI. These processes may be further exemplifiedas processes for preparing hybrid broccoli seed or plants, wherein afirst broccoli plant is crossed with a second broccoli plant of adifferent, distinct genotype to provide a hybrid that has, as one of itsparents, a plant of broccoli line BRM-53-3936CMS or broccoli lineBRM-53-5943SI. In these processes, crossing will result in theproduction of seed. The seed production occurs regardless of whether theseed is collected or not.

In one embodiment of the invention, the first step in “crossing”comprises planting seeds of a first and second parent broccoli plant,often in proximity so that pollination will occur for example, mediatedby insect vectors. Alternatively, pollen can be transferred manually.Where the plant is self-pollinated, pollination may occur without theneed for direct human intervention other than plant cultivation.

A second step may comprise cultivating or growing the seeds of first andsecond parent broccoli plants into plants that bear flowers. A thirdstep may comprise preventing self-pollination of the plants, such as byemasculating the flowers (i.e., killing or removing the pollen).

A fourth step for a hybrid cross may comprise cross-pollination betweenthe first and second parent broccoli plants. Yet another step comprisesharvesting the seeds from at least one of the parent broccoli plants.The harvested seed can be grown to produce a broccoli plant or hybridbroccoli plant.

The present invention also provides the broccoli seeds and plantsproduced by a process that comprises crossing a first parent broccoliplant with a second parent broccoli plant, wherein at least one of thefirst or second parent broccoli plants is a plant of broccoli hybrid RX05990035 and/or broccoli lines BRM-53-3936CMS and BRM-53-5943SI. In oneembodiment of the invention, broccoli seed and plants produced by theprocess are first generation (F₁) hybrid broccoli seed and plantsproduced by crossing a plant in accordance with the invention withanother, distinct plant. The present invention further contemplatesplant parts of such an F₁ hybrid broccoli plant, and methods of usethereof. Therefore, certain exemplary embodiments of the inventionprovide an F₁ hybrid broccoli plant and seed thereof.

In still yet another aspect, the present invention provides a method ofproducing a plant derived from hybrid RX 05990035 and/or broccoli linesBRM-53-3936CMS and BRM-53-5943SI, the method comprising the steps of:(a) preparing a progeny plant derived from hybrid RX 05990035 and/orbroccoli lines BRM-53-3936CMS and BRM-53-5943SI, wherein said preparingcomprises crossing a plant of the hybrid RX 05990035 and/or broccolilines BRM-53-3936CMS and BRM-53-5943SI with a second plant; and (b)crossing the progeny plant with itself or a second plant to produce aseed of a progeny plant of a subsequent generation. In furtherembodiments, the method may additionally comprise: (c) growing a progenyplant of a subsequent generation from said seed of a progeny plant of asubsequent generation and crossing the progeny plant of a subsequentgeneration with itself or a second plant; and repeating the steps for anadditional 3-10 generations to produce a plant derived from hybrid RX05990035 and/or broccoli lines BRM-53-3936CMS and BRM-53-5943SI. Theplant derived from hybrid RX 05990035 and/or broccoli linesBRM-53-3936CMS and BRM-53-5943SI may be an inbred line, and theaforementioned repeated crossing steps may be defined as comprisingsufficient inbreeding to produce the inbred line. In the method, it maybe desirable to select particular plants resulting from step (c) forcontinued crossing according to steps (b) and (c). By selecting plantshaving one or more desirable traits, a plant derived from hybrid RX05990035 and/or broccoli lines BRM-53-3936CMS and BRM-53-5943SI isobtained which possesses some of the desirable traits of the line/hybridas well as potentially other selected traits.

In certain embodiments, the present invention provides a method ofproducing food or feed comprising: (a) obtaining a plant of broccolihybrid RX 05990035 and/or broccoli lines BRM-53-3936CMS andBRM-53-5943SI, wherein the plant has been cultivated to maturity, and(b) collecting tissue of the plant.

In still yet another aspect of the invention, the genetic complement ofbroccoli hybrid RX 05990035 and/or broccoli lines BRM-53-3936CMS andBRM-53-5943SI is provided. The phrase “genetic complement” is used torefer to the aggregate of nucleotide sequences, the expression of whichsequences defines the phenotype of, in the present case, a broccoliplant, or a cell or tissue of that plant. A genetic complement thusrepresents the genetic makeup of a cell, tissue or plant, and a hybridgenetic complement represents the genetic make up of a hybrid cell,tissue or plant. The invention thus provides broccoli plant cells thathave a genetic complement in accordance with the broccoli plant cellsdisclosed herein, and plants, seeds and plants containing such cells.

Plant genetic complements may be assessed by genetic marker profiles,and by the expression of phenotypic traits that are characteristic ofthe expression of the genetic complement, e.g., isozyme typing profiles.It is understood that hybrid RX 05990035 and/or broccoli linesBRM-53-3936CMS and BRM-53-5943SI could be identified by any of the manywell known techniques such as, for example, Simple Sequence LengthPolymorphisms (SSLPs) (Williams et al., Nucleic Acids Res., 18:6531-6535, 1990), Randomly Amplified Polymorphic DNAs (RAPDs), DNAAmplification Fingerprinting (DAF), Sequence Characterized AmplifiedRegions (SCARs), Arbitrary Primed Polymerase Chain Reaction (AP-PCR),Amplified Fragment Length Polymorphisms (AFLPs) (EP 534 858,specifically incorporated herein by reference in its entirety), andSingle Nucleotide Polymorphisms (SNPs) (Wang et al., Science,280:1077-1082, 1998).

In still yet another aspect, the present invention provides hybridgenetic complements, as represented by broccoli plant cells, tissues,plants, and seeds, formed by the combination of a haploid geneticcomplement of a broccoli plant of the invention with a haploid geneticcomplement of a second broccoli plant, preferably, another, distinctbroccoli plant. In another aspect, the present invention provides abroccoli plant regenerated from a tissue culture that comprises a hybridgenetic complement of this invention.

In still yet another aspect, the invention provides a method ofdetermining the genotype of a plant of broccoli hybrid RX 05990035and/or broccoli lines BRM-53-3936CMS and BRM-53-5943SI comprisingdetecting in the genome of the plant at least a first polymorphism. Themethod may, in certain embodiments, comprise detecting a plurality ofpolymorphisms in the genome of the plant. The method may furthercomprise storing the results of the step of detecting the plurality ofpolymorphisms on a computer readable medium. The invention furtherprovides a computer readable medium produced by such a method.

Any embodiment discussed herein with respect to one aspect of theinvention applies to other aspects of the invention as well, unlessspecifically noted.

The term “about” is used to indicate that a value includes the standarddeviation of the mean for the device or method being employed todetermine the value. The use of the term “or” in the claims is used tomean “and/or” unless explicitly indicated to refer to alternatives onlyor the alternatives are mutually exclusive. When used in conjunctionwith the word “comprising” or other open language in the claims, thewords “a” and “an” denote “one or more,” unless specifically notedotherwise. The terms “comprise,” “have” and “include” are open-endedlinking verbs. Any forms or tenses of one or more of these verbs, suchas “comprises,” “comprising,” “has,” “having,” “includes” and“including,” are also open-ended. For example, any method that“comprises,” “has” or “includes” one or more steps is not limited topossessing only those one or more steps and also covers other unlistedsteps. Similarly, any plant that “comprises,” “has” or “includes” one ormore traits is not limited to possessing only those one or more traitsand covers other unlisted traits.

Other objects, features and advantages of the present invention willbecome apparent from the following detailed description. It should beunderstood, however, that the detailed description and any specificexamples provided, while indicating specific embodiments of theinvention, are given by way of illustration only, since various changesand modifications within the spirit and scope of the invention willbecome apparent to those skilled in the art from this detaileddescription.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1: Breeding history for parent line BRM-53-3936CMS, also known asBRM53-3936CMS.

FIG. 2: Breeding history for parent line BRM-53-5943SI, also known asBRM 53-5943SI.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides methods and compositions relating to plants,seeds and derivatives of broccoli hybrid RX 05990035 and/or broccolilines BRM-53-3936CMS and BRM-53-5943SI. The hybrid RX 05990035 wasproduced by the cross of parent lines BRM-53-3936CMS and BRM-53-5943SI.The parent lines show uniformity and stability within the limits ofenvironmental influence. By crossing the parent lines, uniform plants ofhybrid RX 05990035 can be obtained.

Hybrid RX 05990035 does not have a Raised Head, but has clean stem forease of harvest. Hybrid RX 05990035 matures relatively uniform so thatless harvest cuts are required compared to standard broccoli varieties.In uniform conditions even single (machine) harvest is an option.

A. Origin and Breeding History of Broccoli Hybrid RX 05990035

The parents of hybrid RX 05990035 are BRM-53-3936CMS and BRM-53-5943SI.

-   -   B. Physiological and Morphological Characteristics of Broccoli        Hybrid RX 05990035, Broccoli Line BRM-53-3936CMS and Broccoli        Line BRM-53-5943SI

In accordance with one aspect of the present invention, there isprovided a plant having the physiological and morphologicalcharacteristics of broccoli hybrid RX 05990035 and the parent linesthereof. A description of the physiological and morphologicalcharacteristics of such plants is presented in Tables 1-3.

TABLE 1 Physiological and Morphological Characteristics of Hybrid RX05990035 Characteristic RX 05990035 Sibsey A Region of Adaptation NWEurope NW Europe B Maturity: spring planted days from direct seeding tono direct seeding no direct seeding 50% harvest days from transplantingto 50% 60 54 harvest transplant date 10-May-10 10-May-10 length ofharvest period in days  7  7 first harvest date 1-Jul-10 28-June-10 lastharvest date 8-Jul-10 5-Jul-10 Maturity: fall planted days from directseeding to no direct seeding no direct seeding 50% harvest days fromtransplanting to 50% 61 51 harvest transplant date 19-Jul-11 19-Jul-11length of harvest period in days  4  4 first harvest date 18-Sept-114-Sept-11 last harvest date 22-Sep-11 8-Sep-11 time of harvest maturity(50% early very early of plants) (Galaxy, Packman, (Earlyman, Primor)Scorpio) time of beginning of flowering medium early (50% of plants withat least (Coaster, Cruiser) (Clipper, Southern 10% flowers) Comet) CSeedling cotyledon color medium green medium green RHS Color Chart valuefor 147B 137B seedling cotyledon color cotyledon anthocyanin weak weakhypocotyl anthocyanin strong strong D Plant plant height in centimeters60.8 cm 52.1 cm (from soil line to top of head) head height incentimeters 46.5 cm 56.2 cm (from soil line to top of head) height (atharvest maturity) tall medium (Citation) (Coaster) number of stems oneone (Ramoso Calabrese, (Ramoso Calabrese, Shogun) Shogun) branches fewmany habit compact compact market class fresh market fresh market lifecycle annual annual type of variety first generation hybrid firstgeneration hybrid E Leaves outer leaves: number of leaves 14 15 perplant (at harvest) outer leaves: width (at midpoint 21.5 cm 17.8 cm ofplant including petiole) leaf: width medium narrow (Buccaneer, GreenBelt) (Arcadia, Brigadeer) outer leaves: length (at 47.4 cm 36.7 cmmidpoint of plant including petiole) leaf: length (including petiole)medium short (Brigadeer, Sumosun) (Dandy Early, Emperor) outer leaves:petiole length 20.5 cm 15.0 cm petiole: length medium short (Emperor,Ramoso (High Sierra, Calabrese) Padovano) outer leaves: leaf ratio - 2:12:1 length/width outer leaves: leaf attachment petiolate petiolate outerleaves: wax presence weak intermediate leaf: number of lobes mediummedium (Coaster, Topper) (Coaster, Topper) outer leaves: foliage color(with grey-green grey-green wax, if present) outer leaves: foliage color(with 189A 189A wax, if present; RHS Color Chart value) leaf blade:color grey green grey green (Bishop) (Bishop) leaf blade: intensity ofcolor medium medium leaf blade: anthocyanin absent absent coloration(Claudia, Embassy) (Claudia, Embassy) leaf blade: undulation of mediummargin (Citation) leaf blade: dentation of margin weak weak (Galaxy)(Galaxy) outer leaves: leaf shape elliptic elliptic outer leaves: leafbase blunt blunt outer leaves: leaf apex blunt blunt outer leaves: leafmargins slightly wavy straight outer leaves: leaf veins intermediatethin outer leaves: midrib slightly raised not raised leaf blade:blistering weak absent or very weak (Coaster, Gem) (Buccaneer, Colibri)outer leaves: attitude (leaf semi-erect semi-erect angle from ground)(35-55 degrees) (35-55 degrees) leaf: attitude (at beginning ofsemi-erect semi-erect head formation) (Arcadia, Asti, Civet, Claudia)outer leaves: torsion of leaf tip weak weak outer leaves: profile ofupper planar convex side of leaf F Head length of branching at basemedium medium (excluding stem) (Capitol, Green Duke, (Capitol, GreenDuke, Perseus) Perseus) diameter (at widest point) 16.5 cm 12.8 cm (atmarket maturity) depth (at market maturity)  9.7 cm  8.5 cm weight,market trimmed (at 394.0 gm  161.0 gm  market maturity) color (at marketmaturity) blue-green grey-green (Buccaneer) (Brigadeer, Galaxy)intensity of color medium medium RHS Color Chart value for 133B N198Bhead color anthocyanin coloration present absent (Brigadeer, Shogun,(Early White Viola) Sprouting) intensity of anthocyanin very weakcoloration shape (at market maturity) transverse elliptic narrowtransverse elliptic (Citation, Scorpio, Zeus) (Buccaneer, Futura) domeshape (at market domed domed maturity) size (at market maturity) mediumsmall (Dundee, Early Man) (Orbit, Scorpio) compactness/firmness (atmedium medium market maturity) (Late Corona) (Late Corona) surfaceknobbling (at market medium fine maturity) (Southern Comet) (Apollo,Brigadeer) texture fine fine (Auriga, Bishop, Green (Auriga, Bishop,Green Top) Top) bead size (at market maturity) medium small flower buds(at market even in size even in size maturity) anthocyanin coloration ofleaf absent absent axils (at market maturity) anthocyanin coloration ofleaf absent absent veins (at market maturity) anthocyanin coloration ofleaf absent absent blade (at market maturity) anthocyanin coloration ofabsent absent entire plant (at market maturity) anthocyanin colorationof leaf absent absent petiole (at market maturity) (Claudia, Embassy)(Claudia, Embassy) color of head leaves (at market green green maturity)RHS color chart value for the 189B 189A color of head leaves bractsabsent absent (Gem, Orion) (Gem, Orion) secondary heads (at marketcombination combination maturity) **choice for UPOV TG only prominenceof secondary heads intermediate intermediate (at market maturity)(Citation) (Citation) number of secondary heads (at  3  3 marketmaturity) G Flower Color yellow yellow (Brigadeer, Orion) (Brigadeer,Orion) intensity of yellow color light dark (Brigadeer) (Gem, Orion) RHSColor Chart value for 4C 4A flower color stalk color green green RHSColor Chart value for 138B 138B flower stalk color male sterilitypresent present (Chevalier, Montop) (Chevalier, Montop) *These aretypical values. Values may vary due to environment. Other values thatare substantially equivalent are also within the scope of the invention.

TABLE 2 Physiological and Morphological Characteristics of LineBRM-53-3936CMS Characteristic BRM-53-3936CMS Sibsey A Region ofAdaptation NW Europe NW Europe B Maturity: spring planted days fromdirect seeding to 50% no direct seeding no direct seeding harvest daysfrom transplanting to 50% 72 54 harvest transplant date 10-May-1010-May-10 length of harvest period in days 11  5 first harvest date17-Jul-10 1-Jul-10 last harvest date 27-Jul-10 5-Jul-10 harvest season(main crop at 50% spring/summer summer harvest) time of harvest maturity(50% of late very early plants) (Colibri, Ginga, Pollux) (Earlyman,Primor) time of beginning of flowering (50% late early of plants with atleast 10% flowers) (Shogun, Viola) (Clipper, Southern Comet) C Seedlingcotyledon color light green medium green RHS Color Chart value forseedling 138B 147B cotyledon color cotyledon anthocyanin absentintermediate hypocotyl anthocyanin absent intermediate D Plant plantheight in centimeters (from soil 66.7 cm 55.0 cm line to top of leaves)head height in centimeters (from soil 42.0 cm 50.0 cm line to top ofhead) height (at harvest maturity) medium medium (Coaster) (Coaster)number of stems one one (Ramoso Calabrese, (Ramoso Calabrese, Shogun)Shogun) branches medium many habit intermediate compact market classfresh market fresh market life cycle annual annual type of varietyinbred first generation hybrid E Leaves outer leaves: number of leavesper 18 12 plant (at harvest) outer leaves: width (at midpoint of 22.1 cm18.9 cm plant including petiole) leaf: width medium narrow (Buccaneer,Green Belt) (Arcadia, Brigadeer) outer leaves: length (at midpoint of43.7 cm 39.1 cm plant including petiole) leaf: length (includingpetiole) medium short (Brigadeer, Sumosun) (Dandy Early, Emperor) outerleaves: petiole length 21.1 cm 16.0 cm petiole: length medium short(Emperor, Ramoso (High Sierra, Padovano) Calabrese) outer leaves: leafratio - length/width 2:1 2:1 outer leaves: leaf attachment petiolatepetiolate outer leaves: wax presence weak intermediate leaf: number oflobes medium medium (Coaster, Topper) (Coaster, Topper) outer leaves:foliage color (with wax, blue-green grey-green if present) outer leaves:foliage color (with wax, N189B 189A if present; RHS Color Chart value)leaf blade: color blue green grey green (Citation, Esquire, (Bishop)Symphony) leaf blade: intensity of color medium medium leaf blade:anthocyanin coloration absent absent (Claudia, Embassy) (Claudia,Embassy) leaf blade: undulation of margin strong strong (Aikido,Marathon, (Aikido, Marathon, Samurai) Samurai) leaf blade: dentation ofmargin weak weak (Galaxy) (Galaxy) outer leaves: leaf shape broadelliptic elliptic outer leaves: leaf base blunt blunt outer leaves: leafapex blunt blunt outer leaves: leaf margins very wavy very wavy outerleaves: leaf veins thin thin outer leaves: midrib not raised not raisedleaf blade: blistering medium absent or very weak (Medium Late 145,Skiff) (Buccaneer, Colibri) outer leaves: attitude (leaf angle fromsemi-erect semi-erect ground) (35-55 degrees) (35-55 degrees) leaf:attitude (at beginning of head semi-erect semi-erect formation)(Arcadia, Asti, Civet, Claudia) outer leaves: torsion of leaf tip weakweak outer leaves: profile of upper side of convex convex leaf F Headlength of branching at base (excluding medium medium stem) (Capitol,Green Duke, (Capitol, Green Duke, Perseus) Perseus) diameter (at widestpoint) (at market 12.9 cm 13.7 cm maturity) depth (at market maturity)11.7 cm 10.9 cm weight (market trimmed) (at market 225.6 gm 182.7 gmmaturity) color (at market maturity) light green grey-green (Brigadeer,Galaxy) intensity of color medium medium RHS Color Chart value for headcolor 144B N189B anthocyanin coloration absent absent (Early WhiteSprouting) (Early White Sprouting) shape (at market maturity) transversebroad elliptic transverse elliptic (Admiral, Corvet) (Buccaneer, Futura)dome shape (at market maturity) domed domed size (at market maturity)small small (Orbit, Scorpio) (Orbit, Scorpio) compactness/firmness (atmarket medium medium maturity) (Late Corona) (Late Corona) surfaceknobbling (at market maturity) medium fine (Southern Comet) (Apollo,Brigadeer) texture very fine fine (Viola) (Auriga, Bishop, Green Top)bead size (at market maturity) small small flower buds (at marketmaturity) even in size even in size anthocyanin coloration of leaf axils(at absent absent market maturity) anthocyanin coloration of leaf veinsabsent absent (at market maturity) anthocyanin coloration of leaf bladeabsent absent (at market maturity) anthocyanin coloration of entireplant absent absent (at market maturity) anthocyanin coloration of leafpetiole absent absent (at market maturity) (Claudia, Embassy) (Claudia,Embassy) color of head leaves (at market green green maturity) RHS ColorChart value for the color 189A 189A of head leaves bracts absent absent(Gem, Orion) (Gem, Orion) secondary heads (at market maturity)combination combination **choice for UPOV TG only prominence ofsecondary heads (at intermediate intermediate market maturity)(Citation) (Citation) number of secondary heads (at market  3  4maturity) G Flower Color yellow yellow (Brigadeer, Orion) (Brigadeer,Orion) intensity of yellow color light dark (Brigadeer) (Gem, Orion) RHSColor Chart value for flower 4C 4A color stalk color green green RHSColor Chart value for flower 138A 138B stalk color male sterilitypresent present (Chevalier, Montop) (Chevalier, Montop) *These aretypical values. Values may vary due to environment. Other values thatare substantially equivalent are also within the scope of the invention.

TABLE 3 Physiological and Morphological Characteristics of LineBRM-53-5943SI Characteristic BRM-53-5943SI BRM 53-3924SI A Maturity:fall planted days from transplanting to 50% 60 58 harvest transplantdate 3-Jul-12 3-Jul-12 length of harvest period in days 9 6 firstharvest date 28-Aug-12 28-Aug-12 last harvest date 6-Sep-12 3-Sep-12harvest season (main crop at fall fall 50% harvest) time of harvestmaturity (50% medium (Sumosun) medium (Sumosun) of plants) time ofbeginning of flowering medium-early medium-early to (50% of plants withat least medium** (Coaster, 10% flowers) Cruiser) B Seedling cotyledoncolor medium green medium green RHS Color Chart value for 147B 137Aseedling cotyledon color cotyledon anthocyanin absent absent hypocotylanthocyanin intermediate intermediate C Plant plant height incentimeters 42.7 cm 37.3 cm from soil line to top of leaves head heightin centimeters from 42.4 cm 40.5 cm soil line to top of leaves height atharvest maturity short short (Packman, Primor) (Packman, Primor) numberof stems one one (Ramoso Calabrese, (Ramoso Calabrese, Shogun) Shogun)branches medium many habit intermediate intermediate market class freshmarket fresh market life cycle annual annual type of variety parent lineparent line D Leaves outer leaves: number of leaves 20 18 per plant (atharvest) outer leaves: width (at midpoint 15.8 cm 16.5 cm of plantincluding petiole) leaf: width narrow narrow (Arcadia, Brigadeer)(Arcadia, Brigadeer) outer leaves: length (at 39.6 cm 31.0 cm midpointof plant including petiole) leaf: length (including petiole) short short(Dandy Early, Emperor) (Dandy Early, Emperor) outer leaves: petiolelength 9.5 cm 9.1 cm petiole: length short (High Sierra, short (HighSierra, Padovano) Padovano) outer leaves: leaf ratio - 2:1 2:1length/width outer leaves: leaf attachment sessile and petiolatepetiolate (both) outer leaves: wax presence intermediate intermediateouter leaves: foliage color (with dark green grey-green wax, if present)outer leaves: foliage color (with N189A N189B wax, if present; RHS ColorChart value) leaf blade: color green grey green (Claudia, Verflor)(Bishop) leaf blade: intensity of color dark leaf blade: anthocyaninpresent absent coloration (Buccaneer, Pascal) (Claudia, Embassy) leafblade: undulation of weak medium margin (Beaufort, Early Pack,(Citation) Laser, Paladin) leaf blade: dentation of margin weak medium(Galaxy) (Buccaneer) outer leaves: leaf shape narrow elliptic ellipticouter leaves: leaf base blunt blunt outer leaves: leaf apex blunt bluntouter leaves: leaf margins straight slightly wavy outer leaves: leafveins thin intermediate outer leaves: midrib slightly raised not raisedleaf blade: blistering absent or very weak absent or very weak(Buccaneer, Colibri) (Buccaneer, Colibri) outer leaves: attitude (leafsemi-erect semi-erect angle from ground) (35-55 degrees) (35-55 degrees)leaf: attitude (at beginning of semi-erect semi-erect head formation)(Arcadia, Asti, Civet, (Arcadia, Asti, Civet, Claudia) Claudia) outerleaves: torsion of leaf tip none none to weak outer leaves: profile ofupper planar convex side of leaf E Head length of branching at basemedium (excluding stem) (Capitol, Green Duke, Perseus) diameter (atwidest point) 12.2 cm 13.2 cm (at market maturity) depth (at marketmaturity) 4.6 cm 3.9 cm weight, market trimmed (at 264.3 gm 321.3 gmmarket maturity) color (at market maturity) grey-green grey-green(Brigadeer, Galaxy) (Brigadeer, Galaxy) RHS Color Chart value for N189B189A head color anthocyanin coloration absent present (Brigadeer, (EarlyWhite Sprouting) Shogun, Viola) shape (at market maturity) transversebroad elliptic transverse broad (Admiral, Corvet) elliptic (Admiral,Corvet) dome shape (at market domed domed maturity) size (at marketmaturity) medium** (Dundee, Early Man) compactness/firmness (at mediummedium market maturity) (Late Corona) (Late Corona) surface knobbling(at market medium medium maturity) (Southern Comet) (Southern Comet)bead size (at market maturity) large medium flower buds (at marketuneven in size (cateye) even in size maturity) anthocyanin coloration ofleaf absent present axils (at market maturity) anthocyanin coloration ofleaf absent absent veins (at market maturity) anthocyanin coloration ofleaf absent absent blade (at market maturity) anthocyanin coloration ofabsent absent entire plant (at market maturity) anthocyanin colorationof leaf absent absent petiole (at market maturity) (Claudia, Embassy)(Claudia, Embassy) color of head leaves (at market green green maturity)color of head leaves (at market N189A 189A maturity) bracts absentabsent (Gem, Orion) (Gem, Orion) secondary heads (at market basal;Combination; present maturity) present (Marathon, Tribute, (Marathon,Tribute, Late Late Purple Sprouting) Purple Sprouting) prominence ofsecondary heads weak intermediate (at market maturity) (Citation) numberof secondary heads (at  0  1.7 market maturity) F Flower color yellowyellow (Brigadeer, Orion) (Brigadeer, Orion) intensity of yellow colormedium (Capitol, Corvet) RHS Color Chart value for 4B 4C flower colorstalk color green green RHS Color Chart value for 138B 138B flower stalkcolor male sterility absent (Marathon) absent (Marathon) *These aretypical values. Values may vary due to environment. Other values thatare substantially equivalent are also within the scope of the invention.

-   -   C. Breeding Broccoli Plants

One aspect of the current invention concerns methods for producing seedof broccoli hybrid RX 05990035 involving crossing broccoli linesBRM-53-3936CMS and BRM-53-5943SI. Alternatively, in other embodiments ofthe invention, hybrid RX 05990035, line BRM-53-3936CMS, or lineBRM-53-5943SI may be crossed with itself or with any second plant. Suchmethods can be used for propagation of hybrid RX 05990035 and/or thebroccoli lines BRM-53-3936CMS and BRM-53-5943SI, or can be used toproduce plants that are derived from hybrid RX 05990035 and/or thebroccoli lines BRM-53-3936CMS and BRM-53-5943SI. Plants derived fromhybrid RX 05990035 and/or the broccoli lines BRM-53-3936CMS andBRM-53-5943SI may be used, in certain embodiments, for the developmentof new broccoli varieties.

The development of new varieties using one or more starting varieties iswell known in the art. In accordance with the invention, novel varietiesmay be created by crossing hybrid RX 05990035 followed by multiplegenerations of breeding according to such well known methods. Newvarieties may be created by crossing with any second plant. In selectingsuch a second plant to cross for the purpose of developing novel lines,it may be desired to choose those plants which either themselves exhibitone or more selected desirable characteristics or which exhibit thedesired characteristic(s) when in hybrid combination. Once initialcrosses have been made, inbreeding and selection take place to producenew varieties. For development of a uniform line, often five or moregenerations of selfing and selection are involved.

Uniform lines of new varieties may also be developed by way ofdouble-haploids. This technique allows the creation of true breedinglines without the need for multiple generations of selfing andselection. In this manner true breeding lines can be produced in aslittle as one generation. Haploid embryos may be produced frommicrospores, pollen, anther cultures, or ovary cultures. The haploidembryos may then be doubled autonomously, or by chemical treatments(e.g. colchicine treatment). Alternatively, haploid embryos may be growninto haploid plants and treated to induce chromosome doubling. In eithercase, fertile homozygous plants are obtained. In accordance with theinvention, any of such techniques may be used in connection with a plantof the invention and progeny thereof to achieve a homozygous line.

Backcrossing can also be used to improve an inbred plant. Backcrossingtransfers a specific desirable trait from one inbred or non-inbredsource to an inbred that lacks that trait. This can be accomplished, forexample, by first crossing a superior inbred (A) (recurrent parent) to adonor inbred (non-recurrent parent), which carries the appropriate locusor loci for the trait in question. The progeny of this cross are thenmated back to the superior recurrent parent (A) followed by selection inthe resultant progeny for the desired trait to be transferred from thenon-recurrent parent. After five or more backcross generations withselection for the desired trait, the progeny have the characteristicbeing transferred, but are like the superior parent for most or almostall other loci. The last backcross generation would be selfed to givepure breeding progeny for the trait being transferred. The plants of thepresent invention are particularly well suited for the development ofnew lines based on the elite nature of the genetic background of theplants. In selecting a second plant to cross with RX 05990035 and/orbroccoli lines BRM-53-3936CMS and BRM-53-5943SI for the purpose ofdeveloping novel broccoli lines, it will typically be preferred tochoose those plants which either themselves exhibit one or more selecteddesirable characteristics or which exhibit the desired characteristic(s)when in hybrid combination. Examples of desirable traits may include, inspecific embodiments, high seed yield, high seed germination, seedlingvigor, high yield, disease tolerance or resistance, and adaptability forsoil and climate conditions. Consumer-driven traits, such as a headshape, nutritional value, and taste are other examples of traits thatmay be incorporated into new lines of broccoli plants developed

-   -   D. Further Embodiments of the Invention

In certain aspects of the invention, plants described herein areprovided modified to include at least a first desired heritable trait.Such plants may, in one embodiment, be developed by a plant breedingtechnique called backcrossing, wherein essentially all of themorphological and physiological characteristics of a variety arerecovered in addition to a genetic locus transferred into the plant viathe backcrossing technique. The term single locus converted plant asused herein refers to those broccoli plants which are developed by aplant breeding technique called backcrossing, wherein essentially all ofthe morphological and physiological characteristics of a variety arerecovered in addition to the single locus transferred into the varietyvia the backcrossing technique. By essentially all of the morphologicaland physiological characteristics, it is meant that the characteristicsof a plant are recovered that are otherwise present when compared in thesame environment, other than an occasional variant trait that mightarise during backcrossing or direct introduction of a transgene.

Backcrossing methods can be used with the present invention to improveor introduce a characteristic into the present variety. The parentalbroccoli plant which contributes the locus for the desiredcharacteristic is termed the nonrecurrent or donor parent. Thisterminology refers to the fact that the nonrecurrent parent is used onetime in the backcross protocol and therefore does not recur. Theparental broccoli plant to which the locus or loci from the nonrecurrentparent are transferred is known as the recurrent parent as it is usedfor several rounds in the backcrossing protocol.

In a typical backcross protocol, the original variety of interest(recurrent parent) is crossed to a second variety (nonrecurrent parent)that carries the single locus of interest to be transferred. Theresulting progeny from this cross are then crossed again to therecurrent parent and the process is repeated until a broccoli plant isobtained wherein essentially all of the morphological and physiologicalcharacteristics of the recurrent parent are recovered in the convertedplant, in addition to the single transferred locus from the nonrecurrentparent.

The selection of a suitable recurrent parent is an important step for asuccessful backcrossing procedure. The goal of a backcross protocol isto alter or substitute a single trait or characteristic in the originalvariety. To accomplish this, a single locus of the recurrent variety ismodified or substituted with the desired locus from the nonrecurrentparent, while retaining essentially all of the rest of the desiredgenetic, and therefore the desired physiological and morphologicalconstitution of the original variety. The choice of the particularnonrecurrent parent will depend on the purpose of the backcross; one ofthe major purposes is to add some commercially desirable trait to theplant. The exact backcrossing protocol will depend on the characteristicor trait being altered and the genetic distance between the recurrentand nonrecurrent parents. Although backcrossing methods are simplifiedwhen the characteristic being transferred is a dominant allele, arecessive allele, or an additive allele (between recessive anddominant), may also be transferred. In this instance it may be necessaryto introduce a test of the progeny to determine if the desiredcharacteristic has been successfully transferred.

In one embodiment, progeny broccoli plants of a backcross in which aplant described herein is the recurrent parent comprise (i) the desiredtrait from the non-recurrent parent and (ii) all of the physiologicaland morphological characteristics of the recurrent parent as determinedat the 5% significance level when grown in the same environmentalconditions.

New varieties can also be developed from more than two parents. Thetechnique, known as modified backcrossing, uses different recurrentparents during the backcrossing. Modified backcrossing may be used toreplace the original recurrent parent with a variety having certain moredesirable characteristics or multiple parents may be used to obtaindifferent desirable characteristics from each.

With the development of molecular markers associated with particulartraits, it is possible to add additional traits into an established germline, such as represented here, with the end result being substantiallythe same base germplasm with the addition of a new trait or traits.Molecular breeding, as described in Moose and Mumm, 2008 (PlantPhysiology, 147: 969-977), for example, and elsewhere, provides amechanism for integrating single or multiple traits or QTL into an eliteline. This molecular breeding-facilitated movement of a trait or traitsinto an elite line may encompass incorporation of a particular genomicfragment associated with a particular trait of interest into the eliteline by the mechanism of identification of the integrated genomicfragment with the use of flanking or associated marker assays. In theembodiment represented here, one, two, three or four genomic loci, forexample, may be integrated into an elite line via this methodology. Whenthis elite line containing the additional loci is further crossed withanother parental elite line to produce hybrid offspring, it is possibleto then incorporate at least eight separate additional loci into thehybrid. These additional loci may confer, for example, such traits as adisease resistance or a fruit quality trait. In one embodiment, eachlocus may confer a separate trait. In another embodiment, loci may needto be homozygous and exist in each parent line to confer a trait in thehybrid. In yet another embodiment, multiple loci may be combined toconfer a single robust phenotype of a desired trait.

Many single locus traits have been identified that are not regularlyselected for in the development of a new inbred but that can be improvedby backcrossing techniques. Single locus traits may or may not betransgenic; examples of these traits include, but are not limited to,herbicide resistance, resistance to bacterial, fungal, or viral disease,insect resistance, modified fatty acid or carbohydrate metabolism, andaltered nutritional quality. These comprise genes generally inheritedthrough the nucleus.

Direct selection may be applied where the single locus acts as adominant trait. For this selection process, the progeny of the initialcross are assayed for viral resistance and/or the presence of thecorresponding gene prior to the backcrossing. Selection eliminates anyplants that do not have the desired gene and resistance trait, and onlythose plants that have the trait are used in the subsequent backcross.This process is then repeated for all additional backcross generations.

Selection of broccoli plants for breeding is not necessarily dependenton the phenotype of a plant and instead can be based on geneticinvestigations. For example, one can utilize a suitable genetic markerwhich is closely genetically linked to a trait of interest. One of thesemarkers can be used to identify the presence or absence of a trait inthe offspring of a particular cross, and can be used in selection ofprogeny for continued breeding. This technique is commonly referred toas marker assisted selection. Any other type of genetic marker or otherassay which is able to identify the relative presence or absence of atrait of interest in a plant can also be useful for breeding purposes.Procedures for marker assisted selection are well known in the art. Suchmethods will be of particular utility in the case of recessive traitsand variable phenotypes, or where conventional assays may be moreexpensive, time consuming or otherwise disadvantageous. Types of geneticmarkers which could be used in accordance with the invention include,but are not necessarily limited to, Simple Sequence Length Polymorphisms(SSLPs) (Williams et al., Nucleic Acids Res., 1 8:6531-6535, 1990),Randomly Amplified Polymorphic DNAs (RAPDs), DNA AmplificationFingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs),Arbitrary Primed Polymerase Chain Reaction (AP-PCR), Amplified FragmentLength Polymorphisms (AFLPs) (EP 534 858, specifically incorporatedherein by reference in its entirety), and Single NucleotidePolymorphisms (SNPs) (Wang et al., Science, 280:1077-1082, 1998).

-   -   E. Plants Derived by Genetic Engineering

Many useful traits that can be introduced by backcrossing, as well asdirectly into a plant, are those which are introduced by genetictransformation techniques. Genetic transformation may therefore be usedto insert a selected transgene into a plant of the invention or may,alternatively, be used for the preparation of transgenes which can beintroduced by backcrossing. Methods for the transformation of plantsthat are well known to those of skill in the art and applicable to manycrop species include, but are not limited to, electroporation,microprojectile bombardment, Agrobacterium-mediated transformation anddirect DNA uptake by protoplasts.

To effect transformation by electroporation, one may employ eitherfriable tissues, such as a suspension culture of cells or embryogeniccallus or alternatively one may transform immature embryos or otherorganized tissue directly. In this technique, one would partiallydegrade the cell walls of the chosen cells by exposing them topectin-degrading enzymes (pectolyases) or mechanically wound tissues ina controlled manner.

An efficient method for delivering transforming DNA segments to plantcells is microprojectile bombardment. In this method, particles arecoated with nucleic acids and delivered into cells by a propellingforce. Exemplary particles include those comprised of tungsten,platinum, and preferably, gold. For the bombardment, cells in suspensionare concentrated on filters or solid culture medium. Alternatively,immature embryos or other target cells may be arranged on solid culturemedium. The cells to be bombarded are positioned at an appropriatedistance below the macroprojectile stopping plate.

An illustrative embodiment of a method for delivering DNA into plantcells by acceleration is the Biolistics Particle Delivery System, whichcan be used to propel particles coated with DNA or cells through ascreen, such as a stainless steel or Nytex screen, onto a surfacecovered with target cells. The screen disperses the particles so thatthey are not delivered to the recipient cells in large aggregates.Microprojectile bombardment techniques are widely applicable, and may beused to transform virtually any plant species.

Agrobacterium-mediated transfer is another widely applicable system forintroducing gene loci into plant cells. An advantage of the technique isthat DNA can be introduced into whole plant tissues, thereby bypassingthe need for regeneration of an intact plant from a protoplast. ModernAgrobacterium transformation vectors are capable of replication in E.coli as well as Agrobacterium, allowing for convenient manipulations(Klee et al., Bio-Technology, 3(7):637-642, 1985). Moreover, recenttechnological advances in vectors for Agrobacterium-mediated genetransfer have improved the arrangement of genes and restriction sites inthe vectors to facilitate the construction of vectors capable ofexpressing various polypeptide coding genes. The vectors described haveconvenient multi-linker regions flanked by a promoter and apolyadenylation site for direct expression of inserted polypeptidecoding genes. Additionally, Agrobacterium containing both armed anddisarmed Ti genes can be used for transformation.

In those plant strains where Agrobacterium-mediated transformation isefficient, it is the method of choice because of the facile and definednature of the gene locus transfer. The use of Agrobacterium-mediatedplant integrating vectors to introduce DNA into plant cells is wellknown in the art (Fraley et al., Bio/Technology, 3:629-635, 1985; U.S.Pat. No. 5,563,055).

Transformation of plant protoplasts also can be achieved using methodsbased on calcium phosphate precipitation, polyethylene glycol treatment,electroporation, and combinations of these treatments (see, e.g.,Potrykus et al., Mol. Gen. Genet., 199:183-188, 1985; Omirulleh et al.,Plant Mol. Biol., 21(3):415-428, 1993; Fromm et al., Nature,312:791-793, 1986; Uchimiya et al., Mol. Gen. Genet., 204:204, 1986;Marcotte et al., Nature, 335:454, 1988). Transformation of plants andexpression of foreign genetic elements is exemplified in Choi et al.(Plant Cell Rep., 13: 344-348, 1994), and Ellul et al. (Theor. Appl.Genet., 107:462-469, 2003).

A number of promoters have utility for plant gene expression for anygene of interest including but not limited to selectable markers,scoreable markers, genes for pest tolerance, disease resistance,nutritional enhancements and any other gene of agronomic interest.Examples of constitutive promoters useful for plant gene expressioninclude, but are not limited to, the cauliflower mosaic virus (CaMV)P-35S promoter, which confers constitutive, high-level expression inmost plant tissues (see, e.g., Odel et al., Nature, 313:810, 1985),including in monocots (see, e.g., Dekeyser et al., Plant Cell, 2:591,1990; Terada and Shimamoto, Mol. Gen. Genet., 220:389, 1990); a tandemlyduplicated version of the CaMV 35S promoter, the enhanced 35S promoter(P-e35S) the nopaline synthase promoter (An et al., Plant Physiol.,88:547, 1988), the octopine synthase promoter (Fromm et al., Plant Cell,1:977, 1989); and the figwort mosaic virus (P-FMV) promoter as describedin U.S. Pat. No. 5,378,619 and an enhanced version of the FMV promoter(P-eFMV) where the promoter sequence of P-FMV is duplicated in tandem,the cauliflower mosaic virus 19S promoter, a sugarcane bacilliform viruspromoter, a commelina yellow mottle virus promoter, and other plant DNAvirus promoters known to express in plant cells.

A variety of plant gene promoters that are regulated in response toenvironmental, hormonal, chemical, and/or developmental signals can alsobe used for expression of an operably linked gene in plant cells,including promoters regulated by (1) heat (Callis et al., PlantPhysiol., 88:965, 1988), (2) light (e.g., pea rbcS-3A promoter,Kuhlemeier et al., Plant Cell, 1:471, 1989; maize rbcS promoter,Schaffner and Sheen, Plant Cell, 3:997, 1991; or chlorophyll a/b-bindingprotein promoter, Simpson et al., EMBO J., 4:2723, 1985), (3) hormones,such as abscisic acid (Marcotte et al., Plant Cell, 1:969, 1989), (4)wounding (e.g., wunl, Siebertz et al., Plant Cell, 1:961, 1989); or (5)chemicals such as methyl jasmonate, salicylic acid, or Safener. It mayalso be advantageous to employ organ-specific promoters (e.g., Roshal etal., EMBO J., 6:1155, 1987; Schernthaner et al., EMBO J., 7:1249, 1988;Bustos et al., Plant Cell, 1:839, 1989).

Exemplary nucleic acids which may be introduced to plants of thisinvention include, for example, DNA sequences or genes from anotherspecies, or even genes or sequences which originate with or are presentin the same species, but are incorporated into recipient cells bygenetic engineering methods rather than classical reproduction orbreeding techniques. However, the term “exogenous” is also intended torefer to genes that are not normally present in the cell beingtransformed, or perhaps simply not present in the form, structure, etc.,as found in the transforming DNA segment or gene, or genes which arenormally present and that one desires to express in a manner thatdiffers from the natural expression pattern, e.g., to over-express.Thus, the term “exogenous” gene or DNA is intended to refer to any geneor DNA segment that is introduced into a recipient cell, regardless ofwhether a similar gene may already be present in such a cell. The typeof DNA included in the exogenous DNA can include DNA which is alreadypresent in the plant cell, DNA from another plant, DNA from a differentorganism, or a DNA generated externally, such as a DNA sequencecontaining an antisense message of a gene, or a DNA sequence encoding asynthetic or modified version of a gene.

Many hundreds if not thousands of different genes are known and couldpotentially be introduced into a broccoli plant according to theinvention. Non-limiting examples of particular genes and correspondingphenotypes one may choose to introduce into a broccoli plant include oneor more genes for insect tolerance, such as a Bacillus thuringiensis(B.t.) gene, pest tolerance such as genes for fungal disease control,herbicide tolerance such as genes conferring glyphosate tolerance, andgenes for quality improvements such as yield, nutritional enhancements,environmental or stress tolerances, or any desirable changes in plantphysiology, growth, development, morphology or plant product(s). Forexample, structural genes would include any gene that confers insecttolerance including but not limited to a Bacillus insect control proteingene as described in WO 99/31248, herein incorporated by reference inits entirety, U.S. Pat. No. 5,689,052, herein incorporated by referencein its entirety, U.S. Pat. Nos. 5,500,365 and 5,880,275, hereinincorporated by reference in their entirety. In another embodiment, thestructural gene can confer tolerance to the herbicide glyphosate asconferred by genes including, but not limited to Agrobacterium strainCP4 glyphosate resistant EPSPS gene (aroA:CP4) as described in U.S. Pat.No. 5,633,435, herein incorporated by reference in its entirety, orglyphosate oxidoreductase gene (GOX) as described in U.S. Pat. No.5,463,175, herein incorporated by reference in its entirety.

Alternatively, the DNA coding sequences can affect these phenotypes byencoding a non-translatable RNA molecule that causes the targetedinhibition of expression of an endogenous gene, for example viaantisense- or cosuppression-mediated mechanisms (see, for example, Birdet al., Biotech. Gen. Engin. Rev., 9:207, 1991). The RNA could also be acatalytic RNA molecule (i.e., a ribozyme) engineered to cleave a desiredendogenous mRNA product (see for example, Gibson and Shillito, Mol.Biotech., 7:125, 1997). Thus, any gene which produces a protein or mRNAwhich expresses a phenotype or morphology change of interest is usefulfor the practice of the present invention.

-   -   F. Definitions

In the description and tables herein, a number of terms are used. Inorder to provide a clear and consistent understanding of thespecification and claims, the following definitions are provided:

Allele: Any of one or more alternative forms of a gene locus, all ofwhich alleles relate to one trait or characteristic. In a diploid cellor organism, the two alleles of a given gene occupy corresponding locion a pair of homologous chromosomes.

Backcrossing: A process in which a breeder repeatedly crosses hybridprogeny, for example a first generation hybrid (F₁), back to one of theparents of the hybrid progeny. Backcrossing can be used to introduce oneor more single locus conversions from one genetic background intoanother.

Crossing: The mating of two parent plants.

Cross-pollination: Fertilization by the union of two gametes fromdifferent plants.

Diploid: A cell or organism having two sets of chromosomes.

Emasculate: The removal of plant male sex organs or the inactivation ofthe organs with a cytoplasmic or nuclear genetic factor or a chemicalagent conferring male sterility.

Enzymes: Molecules which can act as catalysts in biological reactions.

F₁ Hybrid: The first generation progeny of the cross of two nonisogenicplants.

Genotype: The genetic constitution of a cell or organism.

Haploid: A cell or organism having one set of the two sets ofchromosomes in a diploid.

Linkage: A phenomenon wherein alleles on the same chromosome tend tosegregate together more often than expected by chance if theirtransmission was independent.

Marker: A readily detectable phenotype, preferably inherited incodominant fashion (both alleles at a locus in a diploid heterozygoteare readily detectable), with no environmental variance component, i.e.,heritability of 1.

Phenotype: The detectable characteristics of a cell or organism, whichcharacteristics are the manifestation of gene expression.

Quantitative Trait Loci (QTL): Quantitative trait loci (QTL) refer togenetic loci that control to some degree numerically representabletraits that are usually continuously distributed.

Resistance: As used herein, the terms “resistance” and “tolerance” areused interchangeably to describe plants that show no symptoms to aspecified biotic pest, pathogen, abiotic influence or environmentalcondition. These terms are also used to describe plants showing somesymptoms but that are still able to produce marketable product with anacceptable yield. Some plants that are referred to as resistant ortolerant are only so in the sense that they may still produce a crop,even though the plants are stunted and the yield is reduced.

Regeneration: The development of a plant from tissue culture.

Royal Horticultural Society (RHS) color chart value: The RHS color chartis a standardized reference which allows accurate identification of anycolor. A color's designation on the chart describes its hue, brightnessand saturation. A color is precisely named by the RHS color chart byidentifying the group name, sheet number and letter, e.g., Yellow-OrangeGroup 19A or Red Group 41B.

Self-pollination: The transfer of pollen from the anther to the stigmaof the same plant.

Single Locus Converted (Conversion) Plant: Plants which are developed bya plant breeding technique called backcrossing, wherein essentially allof the desired morphological and physiological characteristics of abroccoli variety are recovered in addition to the characteristics of thesingle locus transferred into the variety via the backcrossing techniqueand/or by genetic transformation.

Substantially Equivalent: A characteristic that, when compared, does notshow a statistically significant difference (e.g., p=0.05) from themean.

Tissue Culture: A composition comprising isolated cells of the same or adifferent type or a collection of such cells organized into parts of aplant.

Transgene: A genetic locus comprising a sequence which has beenintroduced into the genome of a broccoli plant by transformation.

-   -   G. Deposit Information

A deposit of broccoli line BRM-53-5943SI, disclosed above and recited inthe claims, has been made with the American Type Culture Collection(ATCC), 10801 University Blvd., Manassas, Va. 20110-2209. The date ofdeposit was Feb. 2, 2012. The accession number for those deposited seedsof broccoli line BRM-53-5943SI is ATCC Accession No. PTA-12484. Uponissuance of a patent, all restrictions upon the deposit will be removed,and the deposit is intended to meet all of the requirements of 37 C.F.R.§1.801-1.809. The deposit will be maintained in the depository for aperiod of 30 years, or 5 years after the last request, or for theeffective life of the patent, whichever is longer, and will be replacedif necessary during that period.

Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity andunderstanding, it will be obvious that certain changes and modificationsmay be practiced within the scope of the invention, as limited only bythe scope of the appended claims.

All references cited herein are hereby expressly incorporated herein byreference.

What is claimed is:
 1. A seed of broccoli line BRM-53-5943SI, a sampleof seed of said line having been deposited under ATCC Accession NumberPTA-12484.
 2. A plant of broccoli line BRM-53-5943SI, a sample of seedof said line having been deposited under ATCC Accession NumberPTA-12484.
 3. A plant part of the plant of claim
 2. 4. The plant part ofclaim 3, wherein said part is selected from the group consisting of aleaf, an ovule, a floret, pollen, a head, or a cell.
 5. A broccoliplant, or a part thereof, having all the physiological and morphologicalcharacteristics of the broccoli plant of claim
 2. 6. A tissue culture ofregenerable cells of broccoli line BRM-53-5943SI, a sample of seed ofsaid line having been deposited under ATCC Accession Number PTA-12484.7. The tissue culture according to claim 6, comprising cells orprotoplasts from a plant part selected from the group consisting ofembryos, meristems, cotyledons, pollen, leaves, anthers, roots, roottips, pistil, flower, seed and stalks.
 8. A broccoli plant regeneratedfrom the tissue culture of claim 6, wherein the regenerated plantexpresses all of the physiological and morphological characteristics ofbroccoli line BRM-53-5943SI, a sample of seed of said line having beendeposited under ATCC Accession Number PTA-12484.
 9. A method ofproducing seed, comprising crossing the plant of claim 2 with itself ora second plant.
 10. The method of claim 9, wherein the plant of broccoliline BRM-53-5943SI is the female parent.
 11. The method of claim 9,wherein the plant of broccoli line BRM-53-5943SI is the male parent. 12.An F1 hybrid seed produced by the method of claim
 9. 13. An F1 hybridplant produced by growing the seed of claim
 12. 14. A plant produced byintroducing a single locus conversion into broccoli line BRM-53-5943SI,or a selfed progeny thereof, wherein the single locus was introducedinto broccoli line BRM-53-5943SI by backcrossing or genetictransformation and wherein a sample of seed of broccoli lineBRM-53-5943SI has been deposited under ATCC Accession Number PTA-12484.15. The plant of claim 12, wherein the single locus conversion comprisesa transgene.
 16. A seed that produces the plant of claim
 14. 17. Theseed of claim 16, wherein the single locus confers a trait selected fromthe group consisting of male sterility, herbicide tolerance, insectresistance, pest resistance, disease resistance, modified fatty acidmetabolism, environmental stress tolerance, modified carbohydratemetabolism and modified protein metabolism.
 18. The seed of claim 16,wherein the herbicide tolerance trait is tolerance to an herbicideselected from the group consisting of glyphosate, sulfonylurea,imidazalinone, dicamba, glufosinate, phenoxy proprionic acid,cycloshexone, triazine, benzonitrile, PPO-inhibitor herbicides andbroxynil.
 19. A method of producing a seed of a progeny plant derivedfrom the broccoli line BRM-53-5943SI, the method comprising crossing aplant of broccoli line BRM-53-5943SI with a broccoli plant of a secondline to produce at least a first seed, wherein, a sample of seed ofbroccoli line BRM-53-5943SI has been deposited under ATCC AccessionNumber PTA-12484.
 20. The method of claim 19, further comprising thesteps of (a) crossing a plant grown from said seed with itself or asecond plant to produce a seed of a progeny plant of a subsequentgeneration; (b) growing a progeny plant of a subsequent generation fromsaid seed and crossing the progeny plant of a subsequent generation withitself or a second plant; and (c) repeating steps (a) and (b) withsufficient inbreeding until a seed of an inbred broccoli plant derivedfrom the broccoli line BRM-53-5943SI is produced.
 21. The method ofclaim 20, further comprising crossing said inbred broccoli plant derivedfrom broccoli line BRM-53-5943SI with a broccoli plant of a differentgenotype to produce seed of a hybrid plant derived from broccoli lineBRM-53-5943SI.
 22. A method of producing food or feed comprisingobtaining the plant of claim 2 or a part thereof and producing said foodor feed therefrom.